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. 2013 Jun 26;14(7):13231–13240. doi: 10.3390/ijms140713231

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© 2013 by the authors; licensee MDPI, Basel, Switzerland

This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/3.0/).

PMC Copyright notice

The expression patterns of miR-7b and miR-7b*. (A) Basal levels of miR-7b and miR-7b* expression in AtT-20 cells were analyzed by qRT-PCR; (B) Design of miRNA artificial targets. Renilla (hRluc) containing miRNA artificial targets were constructed by inserting multiple miRNA binding sites into the 3′ UTR of a hRluc reporter gene driven by a T7 promoter. The figure shows the nucleotide sequence of the miRNA artificial targets: the red letters are a mutated sequence; (C) Luciferase assay using the miRNA artificial target reporter constructs. COS-7 cells were co-transfected with the miR-7b* mimic oligonucleiotide and miRNA artificial target (1. miR-7b, 2. miR-7b*, 3. miR-7b* mutation I, 4. miR-7b* mutation II, 5. miR-338-3p). Renilla luciferase activity was normalized against firefly luciferase activity and fold changes are compared to miR-338-3p. The results shown are the means of three independent transfections (error bars indicate standard deviations).

The expression patterns of miR-7b and miR-7b*. (A) Basal levels of miR-7b and miR-7b* expression in AtT-20 cells were analyzed by qRT-PCR; (B) Design of miRNA artificial targets. Renilla (hRluc) containing miRNA artificial targets were constructed by inserting multiple miRNA binding sites into the 3′ UTR of a hRluc reporter gene driven by a T7 promoter. The figure shows the nucleotide sequence of the miRNA artificial targets: the red letters are a mutated sequence; (C) Luciferase assay using the miRNA artificial target reporter constructs. COS-7 cells were co-transfected with the miR-7b* mimic oligonucleiotide and miRNA artificial target (1. miR-7b, 2. miR-7b*, 3. miR-7b* mutation I, 4. miR-7b* mutation II, 5. miR-338-3p). Renilla luciferase activity was normalized against firefly luciferase activity and fold changes are compared to miR-338-3p. The results shown are the means of three independent transfections (error bars indicate standard deviations).